Cardiovascular disease (CVD) represents the leading determinant of morbidity and mortality in both developed and underdeveloped countries. Studies suggest cardiovascular diseases (CVD) may be preventable by lifestyle modifications, such as stop smoking, exercise and nutrition. Given the consequences, cost and risks associated with CVD and its medical treatment, there is a need for nutritional intervention in order to prevent or reduce the occurrence of this condition. There are a number of synthetic drug formulations available to prevent atherosclerosis disease. Some examples are Statins, Colestipol, Clofibrate, Questran, Gemfibrozil, among others. However, they have high potential for side effects, including muscle pain, nausea, heartburn, hepatic dysfunction and gastrointestinal discomforts.
CVDs are the number 1 cause of death globally: more people die annually from CVDs than from any other cause. Estimates from the World Health Organization show that cardiovascular disease (CVD) accounted for approximately 17.5 million deaths in 2012 (approximately 31% of all global deaths). Of these deaths, an estimated of 7.4 million were due to coronary heart diseases and 6.7 million were due to strokes. In 2015 was estimated that 89.6 million people in Latin America were affected by any CVD.
CVD also represents a major economic burden on health care systems, in terms of direct (e.g., hospitalizations, rehabilitation services, physician visits, prescription drugs) and indirect costs associated with mortality and morbidity. For the year 2015, the first economic analysis was done in Latin America, showing that the CVD costs about 30.9 thousand millions dollars in the region.
A major factor in CVD is atherosclerosis, a process of accumulating plaques in blood vessels wall. This disease, very hard to treat and almost impossible to reverse, is highly preventable. The development of atherosclerosis is a multifactorial process in which endothelial dysfunction, inflammatory response, modified lipids and lipoproteins, and activated platelets all play significant roles in the process.
(−)-Epicatechin research has recently attracted great interest due to its potential health benefit to humans. In recent years, an increasing number of experimental and clinical studies suggests a protective effect of (−)-epicatechin at doses of 1 or 2 mg/kg of body weight against atherogenesis, oxidative stress, inflammation, and endothelial function. Moreover, publications sustained that (−)-Epicatechin may be effective and beneficial in the prevention and treatment of atherosclerosis.
Cacao beans are the fruit seed of the cacao tree (Theobroma cacao), which are found in warm, moist climates in areas about 20° latitude north and south of the equator, between 500 and 2000 meters above sea level.
Diseases negatively impact on world cacao production, causing considerable losses that can become 30% or more of the productive potential. Among the most potentially dangerous of these diseases are frosty pod (moniliasis), caused by Moniliophthora roreri (moniliasis), and witches' broom, caused by Moniliophthora perniciosa. Over the centuries, the use of cacao has evolved to what we now know as chocolate (processed bean in solid or liquid form containing varying percentages of cacao liquor, cacao butter, sugar, and milk). Numerous polyphenolic compounds are present in the cacao, in which flavonoids, anthocyanins and tannins are the major phenols. The compounds of particular interest in the present invention are flavanols, a subclass of flavonoids.
Cacao flavonoids are characterized as catechins (flavan-3-ols) and include the monomelic forms, (−)-epicatechin and (+)-catechin, and the oligomeric form procyanidins (also termed proanthocyanidins), which are polymeric compounds comprising catechin and epicatechin subunits. It has been reported that 60% of the total phenolic compounds in raw cacao beans are flavanol monomers (epicatechin and catechin) and procyanidins oligomers (dimer to decamer).
(−)-Epicatechin is a major component of the polyphenols in cacao beans and it comprises approximately 35% of the total phenolic content in unfermented Cacao beans. A study reported that flavonoid-enriched cacao powder contain 128.9 mg/g of procyanidins, and particularly 19.36 mg/g of (−)-epicatechin.
Suggestions regarding the existence of possible cacao-dependent health benefits are not an innovative concept. In the past, “Theobroma cacao” was frequently used as a medicine for various diseases, but its medical use progressively disappeared. In contrast to this, recent studies have demonstrated a potential and to a certain extent unanticipated and unexpected role of cacao in “promoting health”. In fact, a large body of evidence supports that dietary intake of catechins might exert some beneficial vascular effects, reduce the risk of cardiovascular morbidity and mortality, and contribute to the prevention of other chronic diseases.A considerable number of epidemiological investigations have generated data that support an association between the intake of flavanol containing foods and a decreased risk of diseases, in particular cardiovascular ones. Researchers have evaluated the outcomes of 15 prospective cohort studies, which aimed at investigating the relationship between the intake of flavonoid containing foodstuffs and the risk of cardiovascular disease. Thirteen of these studies provided evidence supporting a positive correlation between the dietary intake of flavanols and cardiovascular health, with a reduction of cardiovascular disease mortality of up to 65%.Evidence indicates that (−)-epicatechin is the main cacao flavanol associated with cardiovascular effects. (−)-Epicatechin counteracts the action of oxidized LDL on endothelial cells, an action considered pivotal for endothelial dysfunction in the pathogenesis of atherosclerosis. Additionally, it improves the vascular function, lowers blood pressure and improves insulin sensitivity. These compounds reportedly act as free radical scavengers and inhibitors of eicosainoid biosynthesis; in model systems, they also reduce low-density lipoprotein oxidation, prevent platelet aggregation and protect the heart from ischemia injury.(−)-Epicatechin in cacao quenches OH 100 times more effectively than mannitol, a typical OH scavenger. According to Norman Hollenberg, professor of medicine at Harvard Medical School, (−)-epicatechin can reduce the risk of four of the major health problems: stroke, heart failure, cancer and diabetes. He studied the Kuna people in Panama, who drink up to 40 cups of cacao a week, and found that the prevalence of the “big four” is less than 10%. He believes that (−)-epicatechin should be considered essential to the diet and thus classed as a vitamin. Interestingly, data from a population-based cohort study of 1,169 patients link chocolate consumption with decreased mortality after myocardial infarction.
Other studies have shown the effects of (−)-epicatechin on myocardial infarct size and left ventricular remodeling after permanent coronary occlusion. The results demonstrated the unique capacity of (−)-epicatechin to confer cardioprotection in the setting of a severe form of myocardial ischemic injury. Protection was sustained over time and preserved left ventricle structure and function. The cardioprotective mechanism(s) of (−)-epicatechin seemed to be unrelated to protein kinase B(AKT) or extracellular signal-related kinase (ERK) activation. Results yield a reduction in scar (infarct) size of approximately 33%.
A study has reported that the activity of (−)-epicatechin in endothelial cells modulates the endothelial nitric oxide synthase (eNOS) in a favorable direction by (i) preventing a proteasome-mediated loss of eNOS protein due to oxidatively modified low density lipoproteins (LDL) with concomitant protection of endothelial cells against oxidized low density lipoprotein mediated cell death, (ii) ameliorating endothelial nitric oxide (NO) production at the posttranslational level. The study concludes that (−)-epicatechin contribute to protect the integrity of endothelial cells not only by scavenging free radicals but also by maintaining endothelial NO synthase. Concluding that improving the function of the eNOS pathway may be effective and beneficial in the prevention and treatment of atherosclerosis.
Tolerance trials for a green tea catechin supplement have been carried out concluding that the optimum dosage to obtain the best health benefits from catechins was of 800 mg. This dosage is tolerated well by subjects undergoing an overnight fast. Other research reported that a consumption of a dose approached 500 mg of flavonoids may be beneficial in patients with atherosclerotic disease.
A ninety-three patients trial, administering 27 g/day of flavonoid-enriched chocolate containing 850 mg of flavan-3-ols and a content of 90 mg of epicatechin resulted in a significant reduction of peripheral insulin resistance and improvements in insulin sensitivity. Concluding that one year intervention with flavan-3-ols and isoflavones improved biomarkers of CVD risk, highlighting the additional benefit of flavonoids to standard drug therapy in managing CVD risk in postmenopausal type 2 diabetic patients.
The study suggested that epicatechin dose may be a key contributor. Doses of 50 mg epicatechin/day reduced systolic and diastolic blood pressure. For fasting glucose and triglycerides, beneficial effects were observed at only the 50-100-mg/day epicatechin dose. These findings support oral administration of pure (−)-epicatechin mimicking acute vascular effects.
Thus, one of the major pitfalls of (−)-epicatechins is that they are chemically unstable. In solution, they readily undergo oxidation, involving the loss of hydrogen atoms, the generation of a semiquinone radical intermediate and the formation of quinoneoxidised products. A number of factors, including oxygen concentration and pH, influence the stability of (−)-epicatechins. The most crucial factor in (−)-epicatechin degradation is pH; it has been shown that the rate of oxidation increases as the pH increases.
During heat treatment, nonenzymatic browning is developed through the Maillard reaction (MR), accompanied by the formation of a variety of MR products (MRPs). This reaction involves not only reducing sugars and amino acids but also carbonyl compounds resulting from lipidoxidation. Together with oxidation, condensation, and complexation of polyphenol compounds and following protein and starch hydrolysis, MR is responsible for the formation of the characteristic brown color, pleasant aroma, and texture of roasted cacao beans. It was established that MR is responsible for the decrease of reduced sugar and amino acid concentrations observed during the roasting of cacao beans. The reaction occurs extensively in food systems and in vivo. (−)-Epicatechins react with Maillard reactants in model systems; two main reaction products are reported, epicatechin-C5 and -C6 sugar fragment adducts and quenched 3-deoxy-2-hexosulose (a key source C6 to C1 sugar fragments) and consequently inhibited Maillard product formation.
(−)-Epicatechins are rapidly absorbed in the human body, however, their duration in plasma is considerably short and their excretion from the body appears to be fast. This instability has been cited as one of the reasons for the poor bioavailability of these compounds. The oral bioavailability of (−)-epicatechins is low, at less than 5%, with most of the catechin degradation believed to occur under the small intestine conditions where the elevated pH and the presence of reactive oxygen species provide favorable conditions for catechin auto-oxidative reactions.
Limited Transport of catechins in the intestine is due to the Multidrug Resistance Proteins (MRP) and P-glycoprotein (PgP,) known for limiting the uptake of catechins. Combined, poor intestinal transport and stability may result in limiting the absorption of catechins following oral consumption.
By additional way of background, U.S. Pat. No. 7,488,503 B1 relates to an encapsulation composition prepared in an extruder with water as a liquid plasticizer to be able to extrude the first polymer, the starch. The composition has a selected component from the group consisting of a sugar, a polyol, a corn syrup solid, and mixtures thereof. The second food polymer is at least one member selected from the group consisting of gum arabic, gum karaya, gum tragacanth, konjac, larch gum, locust bean gum, guar gum, xanthan gum, sodium carboxymethyl cellulose, agar agar, type A gelatin, type B gelatin, and mixtures thereof. The objective of the encapsulation is the flavoring agent.
U.S. Pat. No. 6,475,510 features a method for the preparation of a bite-dispersion tablets with the ability to disperse quickly in the mouth without the aid of water. The process comprises a dry granulation of one or more drugs blended with an excipient, flavors and a combination of a waxy material and phospholipid. The bite-dispersion tablet has an intense sweetener derived from fruit flavonoids for taste-masking.
WO/2008/086400 describes a method to produce a bioenhanced products by dry blending and solvent spray drying. In one embodiment the solubility-enhancing organic material is polymeric. The products describe by the invention include pharmaceuticals, nutraceuticals, cosmetic, and personal care products for man and animal.
US 2013/0046011 teaches a hot-melt extruded composition that includes about a plant-derived phenolic material, one or more edible or bioerodible excipients, a surface active material, an oral absorption enhancer, and one or more pharmaceutical or food grade additives. The composition has been hot-melt extruded at a temperature substantially below the melting point of the plant-derived phenolic material.
WO/2015/099842 relates to a nutritional composition for fortifying a hot beverage, or transform a hot beverage into an enhanced energy drink.
WO/2011/141708 discloses new particles comprising a tetracycline or one of its pharmaceutically acceptable salts and an antioxidant. Methods of encapsulation of a tetracycline or one of its pharmaceutically acceptable salts and an antioxidant could be spray drying or melt extrusion.
US 2007/0077279 features a composition containing at least a polyphenol and polyethylenglycol, to product food, beverages, dietary supplements, feed, pharmaceuticals and personal care products. It describes the use of polyethylenglycol for masking the bitter taste of such polyphenols. The polyphenols are preferably selected from the group consisting of epigallocatechin gallate, resveratrol, hydroxytyrosol, oleuropein, polyphenols present in green tea extracts, catechins, polyphenols present in extracts of red grape skin, polyphenols present in olives and/or olive waste water, and their mixtures.
US 2015/0374019 relates to a formulations containing isomaltulose and a polyphenol. The same isomaltulose is used for masking unwanted taste components, in particular bitter substances in the formulations containing tea extracts.
A few delivery systems have been developed for catechins in recent years. One system is based on biodegradable photocured polyesters, from which the entrapped (−)-epigallocatechin-3-gallate, catechin from tea, was slowly released upon the erosion of the polymers, for preventing Escherichia coli biofilm formation. Another system used a chewing gum for the slow intake of catechins over a chewing period of about 30-40 min. However, little progress has been reported on an oral delivery system, which can sustainably release the entrapped catechins during digestion. The catechin or (−)-Epicatechins can be protected to prevent any degradation by the addition of ascorbic acid. The prior art is silent in that the protection can be achieved by encapsulation in a polymeric system with Hot Melt Extrusion (HME) technology.
Atherosclerotic pathology is very hard to treat and almost impossible to reverse, but is highly preventable. Given the consequences, cost and risks associated there is a need for nutritional interventions in order to prevent the occurrence of this condition.